肿瘤相关成纤维细胞衍生的细胞外囊泡通过 PD-L1/PD-1 表达介导膀胱癌的免疫逃逸。
Cancer-associated Fibroblast-derived Extracellular Vesicles Mediate Immune Escape of Bladder Cancer via PD-L1/PD-1 Expression.
机构信息
Department of Urology, The First Affiliated Hospital of Soochow University, Suzhou, 215006, Jiangsu, P.R. China.
Department of Urology, Zhenjiang Hospital of Chinese Traditional and Western Medicine, Zhenjiang, 212002, Jiangsu, P.R. China.
出版信息
Endocr Metab Immune Disord Drug Targets. 2023;23(11):1410-1420. doi: 10.2174/1871530323666230228124125.
OBJECTIVE
Bladder cancer (BCa) is a malignant urological tumor with a high prevalence and poor prognosis. Extracellular vesicles (EVs) are increasingly becoming current hotspots owing to their involvement in cancer progression. This paper probed into the action of cancer-associated fibroblast-derived EVs (CAF-EVs) in the immune escape of BCa.
METHODS
CAFs were identified by immunofluorescence. EVs were extracted from CAFs via ultracentrifugation and later characterized. BCa cells (T24 cell line) were co-cultured with CD8+ T cells and then treated with CAF-EVs. The uptake of EVs by T24 cells was examined by confocal laser microscopy. T24 cell apoptosis and invasion were assessed using flow cytometry and invasion assay. CD8+ T cell proliferation was evaluated using CFSE staining. The levels of cytokines (IFN-γ, IL-2, and TNF-α) were measured by ELISA. PD-L1 and PD-1 levels were determined utilizing RT-qPCR and flow cytometry. BCa mouse models were established to identify the effect of CAF-EVs on BCa progression in vivo.
RESULTS
CAF-EVs decreased apoptosis and enhanced invasion of T24 cells, reduced proliferation of CD8+ T cells, and diminished levels of IFN-γ, IL-2, and TNF-α secreted by CD8+ T cells. CAF-EVs promoted the immune escape of T24 cells by carrying PD-L1. Downregulation of PDL1 expression in T24 cells or EVs partially counteracted the promotion of CAF-EVs on immune escape by reducing the binding of PD-L1 and PD-1. Additionally, CAF-EVs raised tumor volume and weight, upregulated PD-L1 expression, and weakened CD8+ T cell infiltration in BCa mice.
CONCLUSION
CAF-EVs facilitate the immune escape of BCa by upregulating PD-L1/PD-1.
目的
膀胱癌（BCa）是一种常见且预后不良的恶性泌尿系统肿瘤。由于其参与癌症进展，细胞外囊泡（EVs）越来越成为当前的热点。本文探讨了癌相关成纤维细胞衍生的 EVs（CAF-EVs）在 BCa 免疫逃逸中的作用。
方法
通过免疫荧光鉴定 CAFs。通过超速离心从 CAFs 中提取 EVs，并对其进行特征描述。将 BCa 细胞（T24 细胞系）与 CD8+T 细胞共培养，然后用 CAF-EVs 处理。通过共聚焦激光显微镜检查 T24 细胞对 EVs 的摄取。通过流式细胞术和侵袭试验评估 T24 细胞凋亡和侵袭。使用 CFSE 染色评估 CD8+T 细胞增殖。通过 ELISA 测量细胞因子（IFN-γ、IL-2 和 TNF-α）的水平。通过 RT-qPCR 和流式细胞术确定 PD-L1 和 PD-1 水平。建立 BCa 小鼠模型，以鉴定 CAF-EVs 在体内对 BCa 进展的影响。
结果
CAF-EVs 降低了 T24 细胞的凋亡率，增强了其侵袭能力，降低了 CD8+T 细胞的增殖率，并降低了 CD8+T 细胞分泌的 IFN-γ、IL-2 和 TNF-α的水平。CAF-EVs 通过携带 PD-L1 促进 T24 细胞的免疫逃逸。下调 T24 细胞或 EVs 中的 PDL1 表达部分抵消了 CAF-EVs 通过减少 PD-L1 和 PD-1 的结合对免疫逃逸的促进作用。此外，CAF-EVs 增加了 BCa 小鼠的肿瘤体积和重量，上调了 PD-L1 的表达，并削弱了 CD8+T 细胞在 BCa 中的浸润。
结论
CAF-EVs 通过上调 PD-L1/PD-1 促进 BCa 的免疫逃逸。
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